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Immunofluorescence - FOXP3 antibody (ab10563)
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Immunohistochemistry (Frozen sections) - FOXP3 antibody (ab10563)
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Western blot - FOXP3 antibody (ab10563)
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Product Name
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FOXP3 antibody
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See all FOXP3 antibodies (19)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to FOXP3
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human FOXP3.
(Peptide available as ab14151.)
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Reacts with
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Hu
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Does not react with
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Ms
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Predicted to react
(species key)
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Mcq
(Due to sequence homology)
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Specificity
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This antibody recognises a single clean band in a HEK293 cell lysate that overexpresses human FOXP3. The band is not seen in untransfected cells and is blocked by the immunising peptide. The antibody has not been tested in WB against endogenous FOXP3.
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Tested applications
(see key)
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Flow Cyt, ICC, IHC-Fr, IHC-P, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
Flow Cyt: Use at a concentration of 4 µg/ml.
ICC: Use at a concentration of 2.5 - 5 µg/ml.
IHC-P: Use at an assay dependent dilution (PMID 19196820).
IHC-Fr: Use at a concentration of 0.625 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately ~50 kDa (predicted molecular weight: 47.2 kDa). Can be blocked with FOXP3 peptide (ab14151).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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ICC: U2OS cells
WB: HEK 293 lysate over expressing human FOXP3
IHC-Fr: human tonsil
FACS: purified human CD4+, CD4+CD25- and CD4+CD25+ cells (see review)
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Cellular localization
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Nuclear
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Research areas
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Immunology >> Adaptive Immunity >> Regulatory T Cells
Chromatin and Nuclear Signaling >> Transcription >> Domain Families >> Forkhead Box >> FOXP
Cell Biology >> Cell Cycle >> Cell Division >> Chromatid Cohesion
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Relevance
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FOXP3 (a 431 amino acid protein) is a member of the forkhead/winged-helix family of transcriptional regulators and is highly conserved across mammals. FOXP3 is essential for normal immune homeostasis.
FOXP3 is stably and constitutively expressed at a high level in CD25 + CD4 positive regulatory T cells, at low level in CD4 positive/CD25 negative cells, and is absent in CD4 negative/CD8 positive T cells. FOXP3 may be a master regulatory gene and a more specific marker of regulatory T cells than other T cells.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.30 - 0.50 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Notes
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There is a reported difference in the human and mouse immune systems regarding FOXP3 expression being induced by T cell activation.
In contrast to the murine system, the activation of human CD4+ CD25- T cells through TCR leads to the increased expression of FOXP3 mRNA, the surface expression of CD25 and gain of regulatory function (see references especially Walker et al, 2003). This suggests that FOXP3 in human is not a unique marker for regulatory T cells.
The pattern of staining reported with ab10563 is consistent with the pattern of other anti-FOXP3 antibodies commercially available and we would welcome your opinion regarding the difference in population stained with FOXP3 antibodies; certain users have indeed observed a staining in human CD25- cells as reported in several papers.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this FOXP3 antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for FOXP3 antibody images, references, products related to ab10563 and other tools.
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FOXP3 antibody images:
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 Immunofluorescence - FOXP3 antibody (ab10563)
Immunofluorescent imaging of human cells (U2OS) with ab10563 (5 ug/ml) confirms the specificity of this anti-FOXP3 antibody, which recognises nuclear signal only; with no background cytoplasmic or nucleolar staining. IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour). Primary antibody used at 5ug/ml in 5% milk / 0.2% TWEEN for 1 hour, secondary antibody for 30 minutes. All blocking and incubation steps carried out at 37 degrees C. Nuclei were stained with Hoechst stain. Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UK
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Immunohistochemistry (Frozen sections) - FOXP3 antibody (ab10563)
FoxP3 immunostaining on a frozen section from human tonsil. Original magnification x100. Nuclear staining was observed. Cigdem Atayar
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Western blot - FOXP3 antibody (ab10563)
Lanes 1 - 6 : FOXP3 antibody (ab10563) at 2 µg/ml
Lane 7 :
Lane 1 : 20µg HEK 293 lysate
Lane 2 : 20µg HEK 293 lysate over expressing human FOXP3
Lane 3 : 20µg HEK 293 lysate over expressing mouse FOXP3
Lane 4 : 20µg HEK 293 lysate with FOXP3 peptide (ab14151) at 1 µg/ml
Lane 5 : 20µg HEK 293 lysate over expressing human FOXP3 with FOXP3 peptide (ab14151) at 1 µg/ml
Lane 6 : 20µg HEK 293 lysate over expressing mouse FOXP3 with FOXP3 peptide (ab14151) at 1 µg/ml
Lane 7 : 20µg HEK 293 lysate
Secondary
Goat polyclonal to Rabbit IgG - H&L (HRP) (ab6721) at 0.2 µg/ml
Performed under reducing conditions.
Predicted band size : 47.2 kDa
Observed band size : 47 kDa (why is the actual band size different from the predicted?)
Exposure time : 30 seconds
This antibody recognises a single clean band in a HEK293 cell lysate that overexpresses human FOXP3. The band is not seen in untransfected cells and is blocked by the immunising peptide.
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References for FOXP3 antibody (ab10563)
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This product has been used in: (two most recent references)
Hansenne I et al. Aire and Foxp3 expression in a particular microenvironment for T cell differentiation. Neuroimmunomodulation 16:35-44 (2009). ICC/IF. PubMed: 19077444
Graciolli FG et al. Phosphorus overload and PTH induce aortic expression of Runx2 in experimental uraemia. Nephrol Dial Transplant 24:1416-21 (2009). PubMed: 19075196
See all 18 publication references for this product.
If you publish research using ab10563 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to FOXP3
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FOXP3 antibody - more information
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Customer reviews (feedback) regarding FOXP3 antibody |
Customer FAQs regarding FOXP3 antibody |
Protocols for FOXP3 antibody |
Price and availability of products related to FOXP3 antibody |
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FOXP3 antibody - related products:
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FOXP3 antibody - other tools:
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Contact Abcam with a Technical Enquiry about FOXP3 antibody |
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All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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